In-cell NMR of intrinsically disordered proteins in prokaryotic cells

Methods Mol Biol. 2012:895:19-31. doi: 10.1007/978-1-61779-927-3_2.

Abstract

In-cell NMR, i.e., the acquisition of heteronuclear multidimensional NMR of biomacromolecules inside living cells, is, to our knowledge, the only method for investigating the three-dimensional structure and dynamics of proteins at atomic detail in the intracellular environment. Since the inception of the method, intrinsically disordered proteins have been regarded as particular targets for in-cell NMR, due to their expected sensitivity to the molecular crowding in the intracellular environment. While both prokaryotic and eukaryotic cells can be used as host cells for in-cell NMR, prokaryotic in-cell NMR, particularly employing commonly used protein overexpression systems in Escherichia coli cells, is the most accessible approach. In this chapter we describe general procedures for obtaining in-cell NMR spectra in E. coli cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Culture Techniques
  • Escherichia coli / chemistry*
  • Escherichia coli / metabolism
  • Isotope Labeling
  • Nuclear Magnetic Resonance, Biomolecular*
  • Prokaryotic Cells / metabolism
  • Protein Conformation
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry

Substances

  • Recombinant Proteins